Assessing the Accuracy of a Handheld Real-Time Fluorescence Microscope for Rapid Intraoperative Diagnosis of Prostate Biopsies

Standard hematoxylin and eosin (H&E) staining for prostate cancer diagnosis is time-consuming, delaying clinical workflows and intraoperative decisions. We evaluated EndoSCell (ES), a handheld real-time fluorescence microscope, as an efficient solution. In this single-center study, tissues from 115 patients undergoing transperineal biopsy and 10 undergoing radical prostatectomy (RP) were dual-stained with fluorescein sodium and methylene blue, then scanned with ES. Real-time images interpreted by urologists were compared with standard histopathology. Analyzing 209 biopsy samples, ES demonstrated high diagnostic accuracy with a sensitivity of 0.86 (95% CI, 0.75–0.93) and specificity of 0.97 (95% CI, 0.92–0.99). Receiver operating characteristic (ROC) curve analysis showed an area under the curve (AUC) of 0.912, while Cohen's Kappa indicated substantial agreement (0.839, p < 0.001). The mean scanning time was rapid at 82.71 s. Favorable consistency was also observed in RP specimens. Consequently, ES achieves high concordance with conventional histopathology. Its dual-staining mechanism streamlines clinical workflows, bridging the gap between diagnosis and treatment to enable one-stop management, while showing significant potential for the intraoperative assessment of surgical margins. ● Handheld EndoSCell microscopy allows real-time prostate tissue scans. ● ES imaging yields high diagnostic concordance with H&E (AUC 0.912). ● Extremely fast scanning (median 82.71 s) streamlines intraoperative workflow. ● ES microscopy shows promise for real-time surgical margin evaluation.